RESUMO
BACKGROUND: Transplantation of bone marrow mesenchymal stem cells (MSCs) is believed to be a new modality for periodontal tissue regeneration. However, little is known about the factors that influence the functions of the transplanted MSCs. Periodontal ligament cells may be implicated in regulating MSC functions. METHODS: To examine the effect of humoral factors (HFs) produced by human periodontal ligament (HPL) cells on human MSC (hMSC) gene expression, hMSCs were cocultured separately with HPL cells. The gene expression of the hMSCs was analyzed by microarray. Moreover, the effect of conditioned medium (CM) from cultures of HPL cells (CM-HPL cells) on the proliferation and mineralizing capacity of hMSCs was examined. RESULTS: Thirty-five genes whose expressions were upregulated more than two-fold and 32 genes whose expressions were downregulated more than two-fold were identified in hMSCs cocultured separately with HPL cells. CM-HPL cells prevented calcification in hMSCs cultured with bone morphogenetic protein-2 but not dexamethasone. CM-HPL cells stimulated cell proliferation in hMSCs, whereas CM from culture of human osteoblasts did not influence the proliferation. CONCLUSIONS: To the best of our knowledge, this is the first study on the global gene expression of MSCs cocultured with periodontal ligament cells. A particular humoral factor released from periodontal ligament cells is suggested to affect differentiation and proliferation in MSCs.
Assuntos
Células da Medula Óssea/fisiologia , Calcificação Fisiológica/fisiologia , Células-Tronco Mesenquimais/fisiologia , Ligamento Periodontal/citologia , Adolescente , Adulto , Células da Medula Óssea/efeitos dos fármacos , Proteína Morfogenética Óssea 2/farmacologia , Calcificação Fisiológica/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura , Meios de Cultivo Condicionados , Dexametasona/farmacologia , Regulação para Baixo , Feminino , Expressão Gênica , Glucocorticoides/farmacologia , Humanos , Proteína 1 Inibidora de Diferenciação/análise , Proteínas Inibidoras de Diferenciação/análise , Leptina/análise , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Análise em Microsséries , Proteínas de Neoplasias/análise , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Ligamento Periodontal/efeitos dos fármacos , Ligamento Periodontal/fisiologia , Regulação para CimaRESUMO
BACKGROUND: We have previously shown that cultured human periodontal ligament (HPL) cells produce nerve growth factor (NGF) and express mRNA of tyrosine kinase receptor (trkA), a high-affinity receptor of NGF. These findings suggest that NGF modulates the differentiation and proliferation of the periodontal ligament cells by paracrine and autocrine functions in vivo. Endothelial cells also express NGF and trkA. Therefore, NGF may regulate functions of periodontal ligament cells and endothelial cells during periodontal tissue regeneration. METHODS: Effects of NGF on expressions of bone/cementum-related proteins (osteocalcin [OC], bone sialoprotein [BSP], bone morphogenetic protein [BMP-7], core binding factor alpha [Cbfa-1], and type I collagen), calcification in HPL cells, and proliferation and mRNA expression of vascular endothelial growth factor (VEGF), an endothelial cell mitogen, in human microvascular endothelial cells (HMVECs) were examined. RESULTS: NGF elevated mRNA levels of OC, BSP, BMP-7, Cbfa-1, and type I collagen and enhanced mineral deposition in cultures of HPL cells. Furthermore, NGF stimulated mRNA expressions of VEGF-A and VEGF-B and cell proliferation in HMVEC. CONCLUSION: These findings suggest that the functional regulation of periodontal ligament cells and endothelial cells by NGF might result in the acceleration of periodontal tissue regeneration in vivo.
Assuntos
Células Endoteliais/efeitos dos fármacos , Fator de Crescimento Neural/farmacologia , Ligamento Periodontal/citologia , Proteínas Morfogenéticas Ósseas/metabolismo , Técnicas de Cultura de Células , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo I/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Relação Dose-Resposta a Droga , Células Endoteliais/metabolismo , Humanos , Sialoproteína de Ligação à Integrina , Osteocalcina/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Sialoglicoproteínas/metabolismo , Fatores de TempoRESUMO
The major problem in cell therapy is the possibility of viral or bacterial infection and immune reactions. Therefore, it is expected of culture cells which are intended to be re-implanted with autologous serum rather than conventional bovine serum. Cell therapy with human mesenchymal stem cells (hMSC), differentiating to various cells, is thought to be curative. To culture hMSC with human autologous serum (HAS) and re-implant them for cell therapy, we developed a completely closed bag system separating serum, comparing proliferation and multipotency of hMSC cultured in HAS with those in foetal calf serum (FCS). HAS was simply, safely and efficiently obtained with the developed closed bag system. Cell proliferation of hMSC cultured in HAS was greater than that in FCS. hMSC, exposed to the defined induction medium containing HAS as well as FCS, differentiated into osteoblasts and adipocytes. These findings suggest that HAS obtained with the developed closed bag system is advantageous in a point of decrease in risk of virus or bacterial infection and foreign protein contamination and enhancement of proliferation of hMSC.
